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Anammox Bacterial Diversity in Various Aquatic Ecosystems Based on the Detection of Hydrazine Oxidase Genes (hzoA/hzoB)

  • Environmental Microbiology
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Abstract

Anammox bacteria belonging to the phylum Planctomycetes are responsible for N removal through NH +4 oxidation coupled with NO −2 reduction. Microbial diversity and ecology of anammox bacteria have not yet been fully revealed due to limitations of 16S rRNA analysis. The hydrazine oxidase gene in cluster 1 (hereafter hzoA/hzoB) was suggested as a proper genetic marker due to its high expression and ubiquitous presence in anammox bacteria. We conducted a comparative analysis of 16S rRNA and hzoA/hzoB genes to reveal anammox bacterial diversity and distribution in various aquatic environments. Phylogenetic analyses of 16S rRNA and hzoA/hzoB genes showed the dominance of Scalindua organisms in marine ecosystems, but there was no congruence of 16S rRNA and hzoA/hzoB gene phylogenies among the freshwater anammox bacteria associated with Brocadia sp., Jettenia sp., and Anammoxoglobus sp. Higher diversity of anammox bacteria was revealed based on hzoA/hzoB genes than 16S rRNA genes in the examined environments. Multiple regression analysis showed that salinity had significant influence on differential distribution and diversity of anammox bacteria in different ecosystems. Thus, molecular detection and resulting phylogeny of the hzoA/hzoB gene generated a better understanding of anammox bacterial diversity and their ecological distribution in various aquatic ecosystems.

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Abbreviations

ANAMMOX:

Anaerobic ammonium oxidation

HZO:

Hydrazine oxidase

UCFR:

Upper Cape Fear River

BR:

Black River

CFRE:

Cape Fear River Estuary

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Acknowledgments

This research was supported by NC Sea Grant, the Ahuja Water Quality Fellowship, and NSF (OCE-0851435). We acknowledge Brian Shirey, Jennifer Bagwell, Jessica Lisa, and Daniel Tozour for their contributions as well as Dr. Steve Carini and Dr. Patrick Erwin for editorial discussion. We appreciate Dr. Kartik Chandran and Dr. Costantino Vetriani for providing activated sludge and hydrothermal vent sediment, respectively. We also acknowledge Dr. Michael Mallin, Mr. Matthew McIver, and the cruise members of the Lower Cape Fear River monitoring program for providing sediment samples and measurements of environmental parameters. Finally, we thank Dr. Martin Palmer and Dr. Rachel Mill at the University of Southampton and the members of 2007 JC18 Cruise for the Montserrat samples as well as the Gillings Family Foundation for the travel support.

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Authors and Affiliations

  1. Center for Marine Science, University of North Carolina Wilmington, Wilmington, NC, 28409, USA

    Matthew D. Hirsch & Bongkeun Song

  2. Department of Biology and Marine Biology, University of North Carolina Wilmington, Wilmington, NC, 28403, USA

    Zachery T. Long & Bongkeun Song

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  1. Matthew D. Hirsch
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Correspondence to Bongkeun Song.

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Supplementary Fig. 1

Primers targeting specific amino acid sequence regions of hzoA/hzoB genes. The primers hzoAB1F, hzoAB1R, hzoAB4F, and hzoAB4R were designed in this study. (DOC 31 kb)

Supplementary Table 1

Environmental parameters measured at different sampling sites (PPT 73 kb)

Supplementary Table 2

hzoA/hzoB gene amplification with different PCR primers and conditions (PPT 18 kb)

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Hirsch, M.D., Long, Z.T. & Song, B. Anammox Bacterial Diversity in Various Aquatic Ecosystems Based on the Detection of Hydrazine Oxidase Genes (hzoA/hzoB). Microb Ecol 61, 264–276 (2011). https://doi.org/10.1007/s00248-010-9743-1

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